scholarly journals Light-Inducible Recombinases for Bacterial Optogenetics

2019 ◽  
Author(s):  
Michael B. Sheets ◽  
Wilson W. Wong ◽  
Mary J. Dunlop
Keyword(s):  
Blue Light ◽  
Temporal Dynamics ◽  
Light Sensitivity ◽  
Ambient Light ◽  
Control Of Gene Expression ◽  
Precise Control ◽  
Alternative Protein ◽  
Split Protein ◽  
Spatiotemporal Control ◽  
Effect Of Light

AbstractOptogenetic tools can provide direct and programmable control of gene expression. Light-inducible recombinases, in particular, offer a powerful method for achieving precise spatiotemporal control of DNA modification. However, to-date this technology has been largely limited to eukaryotic systems. Here, we develop optogenetic recombinases for Escherichia coli which activate in response to blue light. Our approach uses a split recombinase coupled with photodimers, where blue light brings the split protein together to form a functional recombinase. We tested both Cre and Flp recombinases, Vivid and Magnet photodimers, and alternative protein split sites in our analysis. The optimal configuration, Opto-Cre-Vvd, exhibits strong blue light-responsive excision and low ambient light sensitivity. For this system we characterize the effect of light intensity and the temporal dynamics of light-induced recombination. These tools expand the microbial optogenetic toolbox, offering the potential for precise control of DNA excision with light-inducible recombinases in bacteria.

scholarly journals Guard cells control hypocotyl elongation through HXK1, HY5, and PIF4

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Gilor Kelly ◽  
Danja Brandsma ◽  
Aiman Egbaria ◽  
Ofer Stein ◽  
Adi Doron-Faigenboim ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Blue Light ◽  
Opposite Effect ◽  
Red Light ◽  
Guard Cells ◽  
Cell Types ◽  
Hypocotyl Elongation ◽  
Sugar Production ◽  
Effect Of Light

AbstractThe hypocotyls of germinating seedlings elongate in a search for light to enable autotrophic sugar production. Upon exposure to light, photoreceptors that are activated by blue and red light halt elongation by preventing the degradation of the hypocotyl-elongation inhibitor HY5 and by inhibiting the activity of the elongation-promoting transcription factors PIFs. The question of how sugar affects hypocotyl elongation and which cell types stimulate and stop that elongation remains unresolved. We found that overexpression of a sugar sensor, Arabidopsis hexokinase 1 (HXK1), in guard cells promotes hypocotyl elongation under white and blue light through PIF4. Furthermore, expression of PIF4 in guard cells is sufficient to promote hypocotyl elongation in the light, while expression of HY5 in guard cells is sufficient to inhibit the elongation of the hy5 mutant and the elongation stimulated by HXK1. HY5 exits the guard cells and inhibits hypocotyl elongation, but is degraded in the dark. We also show that the inhibition of hypocotyl elongation by guard cells’ HY5 involves auto-activation of HY5 expression in other tissues. It appears that guard cells are capable of coordinating hypocotyl elongation and that sugar and HXK1 have the opposite effect of light on hypocotyl elongation, converging at PIF4.

scholarly journals High-performance chemical- and light-inducible recombinases in mammalian cells and mice

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Benjamin H. Weinberg ◽  
Jang Hwan Cho ◽  
Yash Agarwal ◽  
N. T. Hang Pham ◽  
Leidy D. Caraballo ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mammalian Cells ◽  
High Performance ◽  
Genome Engineering ◽  
Genetic Circuits ◽  
Control Of Gene Expression ◽  
Expression Control ◽  
Gene Expression Control ◽  
High Performance Systems ◽  
Spatiotemporal Control

Abstract Site-specific DNA recombinases are important genome engineering tools. Chemical- and light-inducible recombinases, in particular, enable spatiotemporal control of gene expression. However, inducible recombinases are scarce due to the challenge of engineering high performance systems, thus constraining the sophistication of genetic circuits and animal models that can be created. Here we present a library of >20 orthogonal inducible split recombinases that can be activated by small molecules, light and temperature in mammalian cells and mice. Furthermore, we engineer inducible split Cre systems with better performance than existing systems. Using our orthogonal inducible recombinases, we create a genetic switchboard that can independently regulate the expression of 3 different cytokines in the same cell, a tripartite inducible Flp, and a 4-input AND gate. We quantitatively characterize the inducible recombinases for benchmarking their performances, including computation of distinguishability of outputs. This library expands capabilities for multiplexed mammalian gene expression control.

scholarly journals Effect of light and temperature on seed germination in Tibouchina mutabilis (Vell.) Cogn. (Melastomataceae)

2008 ◽  
Vol 8 (2) ◽  
pp. 63-68 ◽  
Author(s):  
Edson Simão ◽  
Massanori Takaki
Keyword(s):  
Seed Germination ◽  
White Light ◽  
Light Sensitivity ◽  
Pioneer Species ◽  
Continuous Darkness ◽  
Effect Of Light

The effect of light and temperature on Tibouchina mutabilis seed germination was analyzed by isothermic incubations in the range of 10 to 40 °C, with 5 °C intervals under both continuous white light (32.85 µmolm-2s-1) and darkness and alternating temperatures (15-20; 15-25; 15-30; 15-35; 20-25; 20-30; 20-35; 25-30; 25-35 and 30-35 °C) under both photoperiod of 12 hours and continuous darkness. The seeds of T. mutabilis need light to trigger the germination and no germination was observed in darkness. The range of optimum temperatures for germination was between 25 to 30 °C and the 20-25 °C alternating temperatures. These results indicate that T. mutabilis behaves as a pioneer species and daily alternating temperatures did not change the light sensitivity of seeds.

scholarly journals Spatiotemporal control of gene expression boundaries using a feedforward loop

2019 ◽  
Author(s):  
Prasad U. Bandodkar ◽  
Hadel Al Asafen ◽  
Gregory T. Reeves
Keyword(s):  
Gene Expression ◽  
Biological Networks ◽  
Target Gene ◽  
Network Motif ◽  
Control Of Gene Expression ◽  
Intermediate Node ◽  
Morphogen Gradients ◽  
Spatially Distributed ◽  
Spatiotemporal Behavior ◽  
Spatiotemporal Control

AbstractA feed forward loop (FFL) is commonly observed in several biological networks. The FFL network motif has been mostly been studied with respect to variation of the input signal in time, with only a few studies of FFL activity in a spatially distributed system such as morphogen-mediated tissue patterning. However, most morphogen gradients also evolve in time. We studied the spatiotemporal behavior of a coherent FFL in two contexts: (1) a generic, oscillating morphogen gradient and (2) the dorsal-ventral patterning of the early Drosophila embryo by a gradient of the NF-κB homolog Dorsal with its early target Twist. In both models, we found features in the dynamics of the intermediate node – phase difference and noise filtering – that were largely independent of the parameterization of the models, and thus were functions of the structure of the FFL itself. In the Dorsal gradient model, we also found that the dynamics of Dorsal require maternal pioneering factor Zelda for proper target gene expression.

scholarly journals A Simple and Low-Cost Strategy to Improve Conidial Yield and Stress Resistance of Trichoderma guizhouense through Optimizing Illumination Conditions

2022 ◽  
Vol 8 (1) ◽  
pp. 50
Author(s):  
Yifan Li ◽  
Xiya Meng ◽  
Degang Guo ◽  
Jia Gao ◽  
Qiwei Huang ◽  
...  
Keyword(s):  
Blue Light ◽  
Stress Responses ◽  
Stress Resistance ◽  
Low Cost ◽  
Hog Pathway ◽  
Precise Control ◽  
Blue Light Receptor ◽  
Expression Of Genes ◽  
Low Cost Strategy ◽  
Stress Related Genes

Light is perceived by photoreceptors in fungi and further integrated into the stress-activated MAPK HOG pathway, and thereby potentially activates the expression of genes for stress responses. This indicates that the precise control of light conditions can likely improve the conidial yield and stress resistance to guarantee the low cost and long shelf life of Trichoderma-based biocontrol agents and biofertilizers. In this study, effects of wavelengths and intensities of light on conidial yield and stress tolerance to osmotic, oxidative and pH stresses in Trichoderma guizhouense were investigated. We found that 2 μmol photons/(m2 × s) of blue light increased the conidial yield more than 1000 folds as compared to dark condition and simultaneously enhanced conidial stress resistance. The enhanced conidial stress resistance is probably due to the upregulated stress-related genes in blue light, which is under the control of the blue light receptor BLR1 and the MAP kinase HOG1.

scholarly journals A collection of genetic mouse lines and related tools for inducible and reversible intersectional misexpression

2019 ◽  
Author(s):  
Elham Ahmadzadeh ◽  
N. Sumru Bayin ◽  
Xinli Qu ◽  
Aditi Singh ◽  
Linda Madisen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Target Gene ◽  
Genetic Manipulation ◽  
Cell Types ◽  
Biological Processes ◽  
Control Of Gene Expression ◽  
A Cell ◽  
Summary Statement ◽  
Spatiotemporal Control ◽  
Mouse Lines

AbstractThanks to many advances in genetic manipulation, mouse models have become very powerful in their ability to interrogate biological processes. In order to precisely target expression of a gene of interest to particular cell types, intersectional genetic approaches utilizing two promoter/enhancers unique to a cell type are ideal. Within these methodologies, variants that add temporal control of gene expression are the most powerful. We describe the development, validation and application of an intersectional approach that involves three transgenes, requiring the intersection of two promoter/enhancers to target gene expression to precise cell types. Furthermore, the approach utilizes available lines expressing tTA/rTA to control timing of gene expression based on whether doxycycline is absent or present, respectively. We also show that the approach can be extended to other animal models, using chicken embryos. We generated three mouse lines targeted at the Tigre (Igs7) locus with TRE-loxP-tdTomato-loxP upstream of three genes (p21, DTA and Ctgf) and combined them with Cre and tTA/rtTA lines that target expression to the cerebellum and limbs. Our tools will facilitate unraveling biological questions in multiple fields and organisms.Summary statementAhmadzadeh et al. present a collection of four mouse lines and genetic tools for misexpression-mediated manipulation of cellular activity with high spatiotemporal control, in a reversible manner.

scholarly journals Genome expression dynamics reveals parasitism regulatory landscape of the root-knot nematode Meloidogyne incognita and a promoter motif associated with effector genes

2021 ◽  
Author(s):  
Martine Da Rocha ◽  
Caroline Bournaud ◽  
Julie Dazeniere ◽  
Peter Thorpe ◽  
Clement Pellegrin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Meloidogyne Incognita ◽  
Life Stage ◽  
Regulation Of Gene Expression ◽  
Root Knot Nematode ◽  
Root Knot Nematodes ◽  
Control Of Gene Expression ◽  
Precise Control ◽  
Effector Genes ◽  
Spatio Temporal

Root-knot nematodes are the major contributor to the crop losses caused by nematodes. Root-knot nematodes secrete effectors into the plant, derived from two sets of pharyngeal gland cells, to manipulate host physiology and immunity. Successful completion of the life cycle, involving successive molts from egg to adult, covers morphologically and functionally distinct stages and will require precise control of gene expression, including effectors. The details of how root-knot nematodes regulate transcription remain sparse. Here, we report a life stage-specific transcriptome of Meloidogyne incognita. Combined with an available annotated genome, we explore the spatio-temporal regulation of gene expression. We reveal gene expression clusters and predicted functions that accompany the major developmental transitions. Focusing on effectors, we identify a putative cis-regulatory motif associated with expression in the dorsal glands: providing an insight into effector regulation. We combine the presence of this motif with several other criteria to predict a novel set of putative dorsal gland effectors. Finally, we show this motif, and thereby its utility, is broadly conserved across the Meloidogyne genus and termed it Mel-DOG. Taken together, we provide the first genome-wide analysis of spatio-temporal gene expression in a root-knot nematode, and identify a new set of candidate effector genes that will guide future functional analyses.

High-Performance Organic Optocouplers Based on an Organic Photodiode With High Blue Light Sensitivity

2013 ◽  
Vol 34 (10) ◽  
pp. 1295-1297 ◽  
Author(s):  
Dong Li ◽  
Guifang Dong ◽  
Lian Duan ◽  
Deqiang Zhang ◽  
Liduo Wang ◽  
...  
Keyword(s):  
Blue Light ◽  
High Performance ◽  
Light Sensitivity
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